体外分析五年制双语.ppt

体外分析五年制双语,体外分析技术的发展,1959年，BersonReceptorsoflowaffinityrequireligandsofhighconcentration.,可逆性（Reversibility）,TheassociationreactionofreceptorandligandisreversibleIftheconcentrationofligandisdecreased,thecompoundswilldissociate.Bindedligandmaybedisplacedbyanotherligandwhichismoreabundantorhashigheraffinitywiththereceptor.,L+R,L.R+L,L*,L*.R,可识别性（Recognition）,Ligandcanrecognisethespecificsiteofthespecificreceptor;receptorcanalsorecognisethespecificligand.,原理,目的:研究受体亲和力和受体数量基础:受体的可饱和性使一定量的受体只能与一定量的配体发生结合反应.方法:用Scatchard作图,得到反映受体特性的亲和常数（affinityconstant，Ka)和最大结合容量（maximalbindingcapacity，Bmax）,L*+R,L*.R+L*,Receptorcapacity(mol),Knownquantityligand,12345n,receptor,10204080160320,combined,B%,方法,Bmax,NSB,TB,TB:totalbinding;SB:specificbinding;NSB:non-specificbinding,(Initialconcentrationofligand),(Concentrationofreceptor-ligandcomplex),饱和曲线（SaturationCurve）,Scatchardchart,Preparationofreceptor,Takeoutthetissueweneed,andputitin4precooledglassculturecapsule,thengetridofremnantbloodandnon-targettissue;Afterweighing,addsuitablebuffer,andshatteritusingquick-actingdispersor,thenmakethetissuehomogenateusingultrasonicwavecomminuter;Centrifugefor20minunder4,6000rpm，thengetridoftheresiduumoftissueandcells,Centrifugethesupernatantfor30min,15000rpm.Removethesupernatant,andCentrifugeagainfor30min,15000rpm.Removethesupernatantcontainingendogenesisligand,andaddsuitablebuffer.Thepreparationisthencompleted.,Requestoflabeledligand,highaffinitytohomologousreceptor,highspecificityhighspecificactivity(10Ci/mMol),highpurity;hermetization,protectionfromlight,hypothermy,Non-radio-immunityassay,Parttwo,Technique,Enzymeimmunoassay,EIAChemiluminescenceimmunoassay,CLIATime-resolvedfluoroimmunoassay,TrFIA,chemiluminescenceapparatusinBayer,USA,酶免疫分析（Enzymeimmunoassay）,直接化学发光免疫分析(Chemiluminescenceimmuneassay),AKP,luminescence,illuminantsubstrateDioxetane,lightintensitymeasuredbyphotomultiplier,AgorAblabeledbyALP,leadtoAg.Abreaction,Eliminationofnomadicpart,incubation,BECKMANCo.principleofAccessassay,CompetitionmethodSandwichmethod,电化学发光免疫分析（Electrochemiluminesce）,时间分辨荧光免疫分析（Time-resolvedfluoroimmunoassay）,Ab-Eu+,Eu,fluorescence,excitation,铕解离出形成新螯合物,新螯合物在保护微膜内形成,300400500600,0,0.4,0.6,0.8,0,10,20,30,40,wavelength/nm,excitation,emission,0.2,H2O,fluorescencedensity,enhancementfluid,Comparison,ClinicalApplicationofinvitroradioassay,Partthree,Thyroidhormoneandrelevanthormone：TT3,TT4,FT3,FT4,Rt3,thyroid-stimulatinghormone(TSH),thyrotropin-releasinghormone(TRH)Thyroidautoantibodyandantigen：TgAb,TmAb,TsAbandTgantigenParathyroidhormone,calcitonin,osteocalcin,etcInsulin,glucagon,CpeptideSuprarenalglandandrelevanthormone：hydrocortisone,aldosterone,ACTH,etc.,Endocrinesystem,TRH,TSH,inorganiciodine,T3,T4,rT3,ACTH,E2、P、TestosteroneHCG、HPL,CRH,hydrocortisonealdosterone,GnRH,LHFSHPRL,hypothalamus,adenohypophysis,Pituitaryhormone：LH,FSH,PRLGonadhormone：E2,E3,testosterone,progesteroneGenitalhormone：placentallactogen、chorionic-gonadotropinhormone,etc,Gonadsystem,CardiovascularsystemANP,myoglobin,digoxin,feritin,ANGandETNervoussystem-END,GH,NSEHematologicalsystemEP,FE,TX,PG,AlimentarysystemCEA,AFP,HBAg,HBAb,gastrinmotilin,etcUrinarysystem2-MG、UAL、THglycoprotein、SIgA,TumorandothersCEA、CA-50、CA-199、CA-125、